MBRS supported students will participate in research funded by NIH grant R01 AI23523. They will assist in the characterization of cytoskeleton proteins of the protozoan parasite, Entamoeba histolytica. The primary strategy will be microsequencing of peptides obtained from actin rich complex proteins purified by PAGE. The amino acid sequence information will be used to design oligonucleotide primers for the polymerase chain reaction (PCR). Gene specific PCR products will be sequenced and also used to screen DNA libraries to obtain the corresponding genes for sequencing. At each stage the sequence information obtained will be compared to that in computerized databases in order to help identify and functionally characterize the proteins analyzed. Participation in this research will give the MBRS supported students broad exposure to techniques in molecular biology,an opportunity to plan and analyze and trouble shoot experimental procedures, and stimulating intellectual problems to solve. The information gained in this research should contribute to the understanding of the molecular organization of the E. histolytica actin cytoskeleton and the role this dynamic cellular apparatus plays in the pathogenicity of amebiasis. At present one graduate student and one undergraduate student are working in the AI's laboratory under MBRS support.